Stable pegylated interferon formulation
View Patent ↗The present invention relates to lyophilized formulations of pegylated interferon which are prepared using trehalose as a cryoprotectant. The formulations have a low moisture content, which helps stabilize the pegylated interferon during storage of the formulations at room temperature. In addition, methods for preparing these formulations are provided.
1. A solution for preparing a lyophilized powder formulation, the solution comprising a pegylated interferon, one or more cryoprotectant agents, a buffer, a stabilizer, and water, wherein trehalose comprises at least 80%, by weight, of the cryoprotectant agents, and the buffer maintains the pH of the formulation between 4.5 and 7.1.
2. The solution of claim 1 , wherein one of the cryoprotectant agents is sucrose.
3. The solution of claim 1 , wherein trehalose is the sole cryoprotectant agent.
4. The solution of claim 3 , which comprises 90 mg/ml trehalose.
5. The solution of claim 1 , wherein the buffer maintains the pH of the solution between 6.5 and 7.1.
6. The solution of claim 5 , wherein the buffer consists of sodium phosphate dibasic anhydrous and sodium phosphate monobasic monohydrate.
7. The solution of claim 1 , wherein the stabilizer is a polysorbate.
8. The solution of claim 7 , wherein the polysorbate is polysorbate 80.
9. The solution of claim 1 , wherein the pegylated interferon consists of covalent conjugates between PEG molecules and interferon alpha molecules.
10. The solution of claim 9 , wherein the PEG molecules are attached to the interferon molecules with a urethane bond.
11. The solution of claim 10 , wherein at least 15% of the attached PEG molecules are attached to histidine residues of the interferon alpha molecules.
12. The solution of claim 11 , wherein the pegylated interferon alpha is PEG 12000 -interferon alpha-2b.
13. The solution of claim 12 , which consists of between 100 and 300 μg/ml of PEG 12000 -interferon alpha-2b, 90 mg/ml trehalose dihydrate, 1.5 mg/ml sodium phosphate dibasic anhydrous, 1.5 mg/ml sodium phosphate monobasic monohydrate, 0.1 mg/ml polysorbate 80 and water.
14. A lyophilized powder, produced by lyophilization of a solution comprising pegylated interferon, one or more cryoprotectant agents, a buffer, a stabilizer, and water, wherein trehalose comprises at least 80%, by weight, of the cryoprotectants agents, the buffer maintains the pH of the solution between 4.5 and 7.1 and the lyophilized powder has a moisture content of less than 3%.
15. The powder of claim 14 , which has a moisture content of less than 2%.
16. The powder of claim 15 , which has less than 10% unpegylated interferon after storage at 40° C. for 30 days.
17. The powder of claim 15 , which is lyophilized in a cartridge.
18. The powder of claim 17 , wherein the solution consists essentially of between 100 and 300 μg/ml of PEG 12000 -interferon alpha-2b, 90 mg/ml trehalose dihydrate, 1.5 mg/ml sodium phosphate dibasic anhydrous, 1.5 mg/ml sodium phosphate monobasic monohydrate, 0.1 mg/ml polysorbate 80 and water.
19. The powder of claim 18 , wherein the lyophilization is performed using a lyophilization cycle comprising the following steps and condition values:
STEP
CONDITION VALUE
LOADING
Temperature (° C.)
5
Pressure (mTorr)
Ambient
FREEZING
Temperature (° C.)
−50
Pressure (mTorr)
Ambient
Time (hrs)
2
ANNEALING
Temperature (° C.)
−20
Pressure (mTorr)
Ambient
Time (hrs)
6
PRIMARY DRYING
Temperature (° C.)
−20
Pressure (mTorr)
15
Time (hrs)
25
SECONDARY DRYING
Temperature (° C.)
5 to 40
Pressure (mTorr)
15
Time (hrs)
6
UNLOADING
Temperature (° C.)
5
Pressure (mTorr)
Ambient.
20. A process for preparing a lyophilized pegylated interferon formulation, comprising:
providing a solution which comprises a pegylated interferon, one or more cryoprotectants agents, a buffer, a stabilizer, and water, wherein trehalose comprises at least 80%, by weight, of the cryoprotectant agents and the buffer maintains the pH of the formulation between 4.5 and 7.1;
placing the solution into a glass container; and
lyophilizing the solution using a lyophilization cycle that produces a lyophilized powder having a moisture content of less than 3%.
21. The process of claim 20 , wherein the glass container is a vial.
22. The process of claim 20 , wherein the lyophilization cycle comprises:
loading the glass container into a lyophilizer at ambient pressure and a temperature between −55° C. and 5° C.;
incubating the glass container at ambient pressure for 1 to 4 hrs at a temperature between −55° C. and −40° C. to freeze the solution in the glass container;
incubating the glass container containing the frozen solution at ambient pressure for 4 to 8 hours at a temperature between −25° C. and −15° C. to anneal the pegylated interferon in the solution;
drying the frozen and annealed solution under vacuum at a pressure between 8 and 100 milliTorr (mTorr) using a primary drying step and a secondary drying step, wherein the primary drying step is performed for 15 to 35 hrs. at a temperature between −30° C. and −15° C. and the secondary drying step is performed for 5 to 10 hrs. at a temperature starting at 0° C. to 5° C. and ramping up to 40° C. during the secondary drying period; and
unloading the glass container from the lyophilizer at ambient pressure and a temperature of 25° C. or below.
23. The process of claim 22 , wherein the glass container is a cartridge and the lyophilization cycle comprises the following steps and condition values:
STEP
CONDITION VALUE
LOADING
Temperature (° C.)
5
Pressure (mTorr)
Ambient
FREEZING
Temperature (° C.)
−50
Pressure (mTorr)
Ambient
Time (hrs)
2
ANNEALING
Temperature (° C.)
−20
Pressure (mTorr)
Ambient
Time (hrs)
6
PRIMARY DRYING
Temperature (° C.)
−20
Pressure (mTorr)
15
Time (hrs)
25
SECONDARY DRYING
Temperature (° C.)
5 to 40
Pressure (mTorr)
15
Time (hrs)
6
UNLOADING
Temperature (° C.)
5
Pressure (mTorr)
Ambient.
24. A pegylated interferon drug product, which comprises a lyophilized powder in a glass container, wherein the lyophilized powder comprises a pegylated interferon, one or more cryoprotectant agents, a buffer, and a stabilizer, wherein trehalose comprises at least 80%, by weight, of the cryoprotectant agents, and the buffer maintains the pH of the formulation between 4.5 and 7.1.
25. The pegylated interferon drug product of claim 24 , wherein trehalose is the sole cryoprotectant agent.
26. The pegylated interferon drug product of claim 24 , wherein the glass container is a vial.
27. The pegylated interferon drug product of claim 24 , wherein the buffer consists of sodium phosphate dibasic anhydrous and sodium phosphate monobasic monohydrate and the stabilizer is a polysorbate.
28. The pegylated interferon drug product of claim 27 , wherein the polysorbate is polysorbate 80.
29. The pegylated interferon drug product of claim 27 , wherein the pegylated interferon consists of covalent conjugates between PEG molecules and interferon alpha molecules, wherein the PEG molecules are attached to the interferon alpha molecules with a urethane bond.
30. The pegylated interferon drug product of claim 29 , wherein at least 15% of the attached PEG molecules are attached to histidine residues of the interferon alpha molecules.
31. The pegylated interferon drug product of claim 30 , wherein the glass container is a cartridge having first and second chambers,
wherein the lyophilized powder is in the first chamber and the second chamber comprises water.
32. The pegylated interferon drug product of claim 31 , wherein the pegylated interferon alpha is PEG 12000 -interferon alpha-2b.
33. The pegylated interferon drug product of claim 32 , wherein the lyophilized powder consists essentially of 67.5 μg of PEG 12000 -interferon alpha-2b, 60.780 mg trehalose dihydrate, 1.013 mg dibasic sodium phosphate anhydrous, 1.013 mg monobasic sodium phosphate monohydrate, and 0.0675 mg polysorbate 80.
34. The pegylated interferon drug product of claim 32 , wherein the lyophilized powder consists essentially of 108 μg of PEG 12000 -interferon alpha-2b, 60.780mg trehalose dihydrate, 1.013 mg dibasic sodium phosphate anhydrous, 1.013 mg monobasic sodium phosphate monohydrate, and 0.0675 mg polysorbate 80.
35. The pegylated interferon drug product of claim 32 , wherein the lyophilized powder consists essentially of 162 μg of PEG 12000 -interferon alpha-2b, 60.780 mg trehalose dihydrate, 1.013 mg dibasic sodium phosphate anhydrous, 1.013 mg monobasic sodium phosphate monohydrate, and 0.0675 mg polysorbate 80.
36. The pegylated interferon drug product of claim 32 , wherein the lyophilized powder consists essentially of 202.5 μg of PEG 12000 -interferon alpha-2b, 60.780 mg trehalose dihydrate, 1.013 mg dibasic sodium phosphate anhydrous, 1.013 mg monobasic sodium phosphate monohydrate, and 0.0675 mg polysorbate 80.